bHLH Proteins and Neural Development in C. elegans

真核细胞的发育依赖于三个精心安排的过程：分化、形态发生和生长。为了做到这一点，发育中的有机体负责调节这些事件所需基因的空间和时间表达。碱性螺旋-环-螺旋(BHLH)转录因子家族通常通过依赖于对环境提供的感觉信号的适当反应的调节级联来调节其中一些基因的表达。约翰逊实验室的成员对了解bHLH家族蛋白在线虫神经发育中的具体作用很感兴趣。在之前由NSF资助的一项研究中，研究生和本科生确定并启动了一项对bHLH基因hlh-17的分子研究。本研究旨在通过两个特定的目的来更清楚地描述hLH-17的表达和生物学活性：(1)通过下拉试验和酵母双杂交系统寻找其他能够与hLH-17相互作用和/或影响hLH-17的蛋白，以及(2)通过定量RT-PCR来确定hLH-17是否影响或受到调节衰老和长寿的基因的影响。该项目将进一步深入了解HLH-17在线虫神经行为中的作用，提供关于HLH-17的哺乳动物同系物在哺乳动物神经发育中的可能作用的信息，并进一步深入了解为数不多的被认为作为负的、非DNA结合的转录调节因子的bHLH蛋白之一。该项目的更广泛影响是，所有的研究都将在摩根州立大学完成，这是一所位于马里兰州巴尔的摩市的历史上的黑人机构，由本科生研究人员和一名硕士研究生(他们中的大多数将是代表不足的少数族裔或女性)完成，他们所有人都将参加当地、国家和国际研究研讨会。此外，约翰逊实验室有着让所有级别(K-12)的学生参与研究活动的悠久传统，通过在当地小学由本科生领导的演示，通过由PI赞助的针对中学生的为期4-6周的实验室实验，以及通过高中/中学生实习，这一传统将继续下去。

Eukaryotic development depends on three carefully orchestrated processes: differentiation, morphogenesis, and growth. In order to do so, the developing organism is responsible for regulating the spatial and temporal expression of the genes that are required for these events. The basic Helix-Loop-Helix (bHLH) transcription factor family regulates the expression of a number of these genes, often through regulatory cascades that rely on the proper responses to sensory signals provided by the environment. Members of the Johnson laboratory are interested in understanding the specific role of the bHLH family of proteins on neural development in the roundworm, Caenorhabditis elegans. In a previous NSF-supported study, graduate and undergraduate students identified and initiated a molecular study of one bHLH gene, hlh-17. The research to take place in the present study is designed to delineate more clearly the expression and biological activity of hlh-17 through two specific aims: (1) identify other bHLH proteins that are capable of interacting with and/or influencing HLH-17 using pull-down assays and the yeast two-hybrid system, and (2) determine if hlh-17 influences, or is influenced by, genes that regulate aging and longevity by using quantitative RT-PCR. This project should provide further insight into the role of HLH-17 in neurological behaviors of C. elegans, provide information on the possible roles of the mammalian homologs of HLH-17 in mammalian neural development, and provide further insight into one of the few bHLH proteins that is believed to act as a negative, non-DNA binding regulator of transcription. The broader impact of the project is that all of the research will be done at Morgan State University, a historically black institution located in Baltimore City, Maryland, by undergraduate student researchers and a Master's degree candidate (most of whom will be underrepresented minorities or women), all of whom will participate in local, national, and international research symposia. In addition, the Johnson laboratory has a long-standing tradition of involving students at all levels (K-12) in the research activities, through undergraduate student-led demonstrations at local elementary schools, through 4-6 week long PI-sponsored laboratory experiments with middle school students, and through high school/middle school student internships, a tradition that will continue with this project.